MHC class II expression and potential antigen-presenting cells in the retina during experimental autoimmune uveitis
Controversy exists regarding which cell types are responsible for autoantigen presentation in the retina during experimental autoimmune uveitis (EAU) development. In this study, we aimed to identify and characterize the retinal resident and infiltrating cells susceptible to express major histocompat...
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Published in | Journal of neuroinflammation Vol. 14; no. 1; p. 136 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
BioMed Central Ltd
18.07.2017
BioMed Central BMC |
Subjects | |
Online Access | Get full text |
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Summary: | Controversy exists regarding which cell types are responsible for autoantigen presentation in the retina during experimental autoimmune uveitis (EAU) development. In this study, we aimed to identify and characterize the retinal resident and infiltrating cells susceptible to express major histocompatibility complex (MHC) class II during EAU.
EAU was induced in C57BL/6 mice by adoptive transfer of autoreactive lymphocytes from IRBP1-20-immunized animals. MHC class II expression was studied by immunostainings on eye cryosections. For flow cytometry (FC) analysis, retinas were dissected and enzymatically digested into single-cell suspensions. Three MHC class II
retinal cell populations were sorted by FC, and their RNA processed for RNA-Seq.
Immunostainings demonstrate strong induction of MHC class II expression in EAU, especially in the inner retina at the level of inflamed vessels, extending to the outer retinal layers and the subretinal space in severely inflamed eyes. Most MHC class II
cells express the hematopoietic marker IBA1. FC quantitative analyses demonstrate that MHC class II induction significantly correlates with disease severity and is associated with upregulation of co-stimulatory molecule expression. In particular, most MHC class II
cells express co-stimulatory molecules during EAU. Further phenotyping identified three MHC class II
retinal cell populations: CD45
CD11b
non-hematopoietic cells with low MHC class II expression and CD45
CD11b
hematopoietic cells with higher MHC class II expression, which can be further separated into Ly6C
and Ly6C
cells, possibly corresponding to infiltrating macrophages and resident microglia. Transcriptome analysis of the three sorted populations leads to a clear sample clustering with some enrichment in macrophage markers and microglial cell markers in Ly6C
and Ly6C
cells, respectively. Functional annotation analysis reveals that both hematopoietic cell populations are more competent in MHC class II-associated antigen presentation and in T cell activation than non-hematopoietic cells.
Our results highlight the potential of cells of hematopoietic origin in local antigen presentation, whatever their Ly6C expression. Our work further provides a first transcriptomic study of MHC class II-expressing retinal cells during EAU and delivers a series of new candidate genes possibly implicated in the pathogenesis of retinal autoimmunity. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1742-2094 1742-2094 |
DOI: | 10.1186/s12974-017-0915-5 |